CN103444535A - Novel method for increasing tissue culture and rapid propagation coefficients of banana - Google Patents
Novel method for increasing tissue culture and rapid propagation coefficients of banana Download PDFInfo
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Abstract
The invention discloses a novel method for increasing tissue culture and rapid propagation coefficients of banana. Transplanted seedlings are obtained through selection and bud induction of explants, differentiation and cultivation of buds, cluster bud toughening, rooting cultivation, seedling hardening and transplanting. On the basis of a conventional method, the novel method has the advantages that a conventional bud suction and cultivation method is improved, so that the number of first generation germination of each bud suction material is 30-40 and is 15-20 times higher than that of the conventional method, and the propagation coefficient is increased to 15-20. The production period of the same number of tissue culture seedlings is shortened by 3-4 months compared with that of the conventional method; the demand on corm is reduced by 15-20 times; the production period is shortened, and the production cost is reduced. The subculture coefficient is low; the number of first generation germination in the conventional method is 2-3, and the number of first generation germination of the novel method is 30-40, so that new bud generations with the same amount are reduced by 3-4 generations; the tissue cultivation seedlings are robust; the aberration rate is low; the operability is high.
Description
Technical field:
The invention belongs to fruit breeding field, be specifically related to a kind of new method that improves banana tissue-culturing rapid propagation coefficient.
Background technology:
Banana occupies important economic status in world's tropical fruit (tree) and Tropical China Production of fruit, is the fourth-largest cereal crops that are only second to paddy rice, wheat and maize, so banana production concern world food safety, regional development and human health.The banana producing region is across more than 130 countries and regions, the whole world.According to the FAO statistics, more than 130 countries in the whole world (area) production banana in 2011, harvest area reaches 5,150,000 hectares, and gross yield reaches 1.065 hundred million tons.China is banana Origin of cultivation ground and big producing country, reaches 40.3 ten thousand hectares to China's banana production area in 2011, and gross yield is 1,071 ten thousand tons, has become the second largest banana production state (FAO, 2013) that global output is only second to India.Banana industry has become China's South Subtropical Area of China agricultural mainstay industry, in hot-zone economy and rural society development, plays an important role.
Fragrant (greatly) any of several broadleaf plants belongs to Musaceae (Musaceae) Musa (Musa), and original banana is divided into wild sharp leaf any of several broadleaf plants (Musa acuminata) and long ridge any of several broadleaf plants (Musa balbisiana).Eat banana raw and be mostly triploid (AAA), cultivation banana (Musa, spp.) is unisexuality triploid (AAA, AAB, or ABB), has the height sterility.
Banana is the large-scale herbaceous monocotyledon of perennial evergreen, and subterranean stem is a thick bulb, root, leaf, flower and breed suction bud used and grow thus.At present banana breeding mainly contains following several method, and a kind of is directly to utilize the suction bud of maternal plant growth to be bred, but the method is owing to producing, to inhale bud speed slower, and reproduction coefficient is very low, has been unwell to modern large-scale commodity production and progressively is eliminated; Also have a kind of it is reported to utilize banana prematurity male flower for the explant rapid propagation in vitro, although its reproduction coefficient is higher, but this method needs first evoked callus, again callus is broken up, operating technology and process are comparatively complicated, difficulty is difficult for more greatly grasping, and the cycle is quite long, is difficult to use in large-scale production.The means that are most widely used at present are to utilize suction gemmule stem manually to carry out the tissue culture technique of numerous bud.The method research and development start from the eighties in last century, have greatly promoted the fast development of China's banana industry, have greatly shortened breeding and promoting of improved seeds, and banana large-scale production is become a reality.But still have at present deficiency and need improved place, the main problem existed is:
1. in conventional method, each bulb sprouting in the first generation is 2-3, and in extensive seedling is produced, annual the needs gathers a large amount of banana bulbs, makes easily band virus, and maternal plant is produced to injury.The explant of cultivating due to tissue is the banana bulb, and bulb grows in soil, easily infects multiple germ, needs strict screening sterilize and pass through virus and detect, and has increased the seedling cost.Any of several broadleaf plants strain bulb collection meeting damages maternal plant frequently in addition, on its output and the certain impact of mass formation.
2. traditional numerous bud technology is because reproduction coefficient is lower, generally only has the 2-3 left and right, for obtaining a large amount of seedlings, needs repeatedly subculture to cultivate, and, along with the increase of subculture number, seedling quality can descend and make aberration rate to raise gradually.Excessively increase the subculture number in large-scale production, can cause a large amount of seedlings inferior to come into the market, infringement banana peasant interests, affect industry development.
3. production cost is higher.Tissue culture seedlings of bananas production is the industry of a labour intensive, and reproduction coefficient is lower, and the production cycle is longer, has consumed a large amount of manpowers and the means of production, has increased production cost.
The deficiency that the above traditional banana group training mode of production exists, started to restrict the development of the quick health of banana industry.Be in particular in: the seedling quality of the too high initiation of subculture coefficient descends; The defect of method of drawing material produces injury to maternal plant, and reproduction coefficient is lower and production cost is improved etc.For deficiency, traditional banana tissue culture and rapid propagation method is improved, guaranteed that the lasting healthy life power of banana seedling industry has become the banana production problem demanding prompt solution.
Summary of the invention:
The objective of the invention is for deficiency of the prior art, a kind of new method that improves banana tissue-culturing rapid propagation coefficient is provided, the method can make the banana bulb first generation number that sprouts reach 30-40, than conventional method, exceeds 15-20 doubly, and reproduction coefficient is increased to the level of 15-20.Produce the group training seedling production cycle of equal number and compare shortening with Traditional Method 3-4 month, and the demand of bulb is reduced to 15-20 doubly, effectively shorten the production cycle, and greatly save production cost.
The new method of raising banana tissue-culturing rapid propagation coefficient of the present invention, is characterized in that, comprises the following steps:
Choosing with bud of a, explant induced: get the suction bud of healthy banana plant base portion, clean, ream and inhale bud top layer and leaf sheath, leave and take the bastem section of inhaling, sterilization treatment, then peel off and inhale bud surface leaf sheath, leave and take the bastem section of inhaling, be placed on inducing culture, 28 ℃ of dark culturing, inhale sprout tuber until grow, and every liter of described inducing culture contains: 6-benzyl aminopurine (6-BA) 3-6mg, methyl α-naphthyl acetate (NAA) 0.1-0.3mg, sucrose 30g, agar 6g, all the other are the MS medium, pH5.6-6.0;
The differentiation of b, bud and cultivation: will inhale the sprout tuber stripping and slicing, be placed in differential medium and carry out the light cultivation, luminous intensity is 1500lx, 28 ℃, to the healthy indefinite bud of differentiation generation, indefinite bud is inoculated on differential medium, in luminous intensity, be 1500lx, 28 ℃ are carried out the subculture cultivation, subculture is cultivated and can be carried out some generations, obtains Multiple Buds, and every liter of described differential medium contains: 6-benzyl aminopurine (6-BA) 3-6mg, methyl α-naphthyl acetate (NAA) 0.1-0.3mg, sucrose 35g, agar 6g, all the other are the MS medium, pH5.6-6.0;
C. the strong bud of Multiple Buds is processed: after Multiple Buds is divided into to little Cong or simple bud, put into strong bud medium, under 28 ℃ of conditions, luminous intensity is 1500lx, grow up to plantlet, every liter of described strong bud medium contains: 6-benzyl aminopurine (6-BA) 0.5-1mg, methyl α-naphthyl acetate (NAA) 0.1mg, sucrose 40g, agar 6g, and all the other are the MS medium, pH5.6-6.0;
D, culture of rootage: choosing is highly the plantlet of 3～4cm, the root induction in root media of transferring, under 28 ℃ of conditions, luminous intensity is 1500lx, cultivate until grow up to Banana Seedlings, every liter of described root media contains methyl α-naphthyl acetate (NAA) 0.1-0.2mg, inositol 50-100mg, sucrose 40g, agar 6g, active carbon 1g, and all the other are the MS medium, pH5.6-6.0;
F, hardening and transplanting: choose well developed root system and healthy and strong Banana Seedlings, take out afterwash root medium, in greenhouse casting bed or Nutrition Soil, after domestication, can be used as and transplant seedling.
Transplant seedling and can be transplanted to field, give suitable rich water quality management and get final product.
Bud top layer and leaf sheath are inhaled in reaming in described step a, leave and take and inhale bastem section, sterilization treatment, peel off and inhale bud surface leaf sheath, leaving and taking suction bastem section is to ream to inhale bud top layer and leaf sheath, leaves and takes the suction bastem section of 5cm, clean with volume fraction 75% alcohol water blend, and use mass fraction 0.1%HgCl
2solution soaks 10 minutes, through aseptic water washing 4-5 time, then blots surface moisture with aseptic filter paper, finally leaves and takes the suction bastem section of 2cm, and peels off layer by layer and inhale the surperficial leaf sheath of bud.
The present invention, than prior art, has following beneficial effect:
1. on the basis of conventional method relatively, tradition is inhaled to bud is processed and cultural method is improved, make each inhale bud material first generation number that sprouts and reach 30-40, exceed 15-20 doubly than conventional method, reproduction coefficient is increased to the level of 15-20.Produce the group training seedling production cycle of equal number and compare shortening with Traditional Method 3-4 month, and the demand of bulb is reduced to 15-20 doubly, effectively save provenance, and shorten the production cycle and save production cost.
2. the subculture number is low: compare in conventional method the first generation and sprout number for 2-3, this new method first generation is sprouted number for 30-40, and the sprouting algebraically of producing equal number reduces 3-4 generation, the group training seedling stalwartness of emerging, and aberration rate is low.Therefore can effectively prevent the bud Quality Down and the high problem of aberration rate that produce because the subculture number is too high.
3. strong operability, compare and utilize banana prematurity male flower to be easier to grasp for explant rapid propagation in vitro method, can be for large-scale production.
Embodiment:
Following examples are to further illustrate of the present invention, rather than limitation of the present invention.
Embodiment 1:
1. material: Brazilian Banana, pick up from Guangdong Academy of Agricultural Sciences's fruit tree research proving ground
2. method
2.1 choosing with bud of explant induced: the suction bud that takes healthy banana plant base portion from field, wash down with running water, and ream and inhale bud top layer leaf sheath with cutter, leave and take the suction bastem section of about 5cm, clean with volume fraction 75% alcohol water blend, and use mass fraction 0.1%HgCl
2solution soaks 10 minutes, through aseptic water washing 4-5 time, with aseptic filter paper, blots surface moisture.Excision is inhaled the bud edge and is left and taken the suction bastem section of about 2cm, peel off layer by layer and inhale bud surface leaf sheath with cutter again, be placed on inducing culture, under 28 ℃ of conditions, dark culturing, after suction bastem section is inoculated in inducing culture 3-4 week, base portion starts to expand, and the rice white growing point, appear in visible significantly callus, after one or two week, growing point becomes obvious healthy budlet, obtain inhaling sprout tuber, each is inhaled sprout tuber number that sprouts and reaches 30～40, than conventional method several 2-3 that sprout, exceeds 15-20 doubly, reproduction coefficient is increased to the level of 15-20; Every liter of described inducing culture contains: 6-benzyl aminopurine (6-BA) 3mg, methyl α-naphthyl acetate (NAA) 0.1mg, sucrose 30g, agar 6g, all the other are the MS medium, pH5.6, and its compound method is that above-mentioned substance is mixed, adjust pH, then high-temperature sterilization is standby.
2.2 the differentiation of bud and cultivation: will inhale sprout tuber and be cut into approximately every 0.5cm size, be placed in differential medium and carry out light cultivation (luminous intensity is 1500lx), 28 ℃, to the healthy indefinite bud of differentiation generation, indefinite bud is inoculated on differential medium, in luminous intensity, is 1500lx, and 28 ℃ are carried out the subculture cultivation, carried out the subculture cultivation once every 25-30 days, subculture obtains Multiple Buds after cultivating; Every liter of described differential medium contains: 6-BA3mg, NAA0.1mg, sucrose 35g, agar 6g, and all the other are the MS medium, pH5.6, its compound method is that above-mentioned substance is mixed, adjust pH, then high-temperature sterilization is standby.
2.3 the strong bud of Multiple Buds is processed: after large Multiple Buds is divided into to little Cong or simple bud, put into strong bud medium, under 28 ℃ of conditions, luminous intensity is 1500lx, and approximately 25 days indefinite buds can grow up to the plantlet of 3-4cm, every liter of described strong bud medium contains: 6-BA0.5mg, NAA0.1mg, sucrose 40g, agar 6g, all the other are the MS medium, pH6.0, and its compound method is that above-mentioned substance is mixed, adjust pH, then high-temperature sterilization is standby.
2.4 culture of rootage: choosing is highly the plantlet of 3-4cm, the root induction in root media of transferring, and under 28 ℃ of conditions, luminous intensity is 1500lx.After 10 days, the seedling base section dissolves the former base of white root, can grow to the 5-7cm Banana Seedlings after 30 days, every liter of described root media contains NAA0.1mg, inositol 50mg, sucrose 40g, agar 6g, active carbon 1g, all the other are the MS medium, pH6.0, its compound method is that above-mentioned substance is mixed, adjust pH, then high-temperature sterilization is standby.
2.5 hardening and transplanting: choose well developed root system and healthy and strong Banana Seedlings, take out afterwash root medium, in greenhouse casting bed or Nutrition Soil, domestication, after 2 months, can be used as and transplants seedling replanting to field, then gives suitable rich water quality management and get final product.
Embodiment 2:
1. material: wide powder dwarf banana, pick up from Guangdong Academy of Agricultural Sciences's white clouds proving ground
2. method
2.1 choosing with bud of explant induced: the suction bud that takes healthy banana plant base portion from field, wash down with running water, and ream and inhale bud top layer leaf sheath with cutter, leave and take the suction bastem section of about 5cm, clean with volume fraction 75% alcohol water blend, and use mass fraction 0.1%HgCl
2solution soaks 10 minutes, through aseptic water washing 4-5 time, with aseptic filter paper, blots surface moisture.Excision is inhaled the bud edge and is left and taken the suction bastem section of about 2cm, peel off layer by layer and inhale bud surface leaf sheath with cutter again, be placed on inducing culture, under 28 ℃ of conditions, dark culturing, inhale bastem section and be inoculated in inducing culture after 3 weeks, base portion starts to expand, visible a small amount of callus and vitrifying tissue, and the rice white growing point appears, after one week, growing point becomes obvious healthy budlet, obtain inhaling sprout tuber, each is inhaled bud fritter number that sprouts and reaches 30～40, than conventional method several 2-3 that sprout, exceeds 15-20 doubly, reproduction coefficient is increased to the level of 15-20; Every liter of described inducing culture contains: 6-benzyl aminopurine (6-BA) 6mg, methyl α-naphthyl acetate (NAA) 0.3mg, sucrose 30g, agar 6g, all the other are the MS medium, pH6.0, and its compound method is that above-mentioned substance is mixed, adjust pH, then high-temperature sterilization is standby.
2.2 the differentiation of bud and cultivation: will inhale sprout tuber and be cut into approximately every 0.5cm size, be placed in differential medium and carry out light cultivation (luminous intensity is 1500lx), 28 ℃, to the healthy indefinite bud of differentiation generation, indefinite bud is inoculated on differential medium, in luminous intensity, is 1500lx, and 28 ℃ are carried out the subculture cultivation, carried out the subculture cultivation once every 25-30 days, subculture obtains Multiple Buds after cultivating; Every liter of described differential medium contains: 6-BA6mg, NAA0.3mg, sucrose 35g, agar 6g, and all the other are the MS medium, pH6.0, its compound method is that above-mentioned substance is mixed, adjust pH, then high-temperature sterilization is standby.
2.3 the strong bud of Multiple Buds is processed: after large Multiple Buds is divided into to little Cong or simple bud, put into strong bud medium, under 28 ℃ of conditions, luminous intensity is 1500lx, and approximately 25 days indefinite buds can grow up to the plantlet of 3-4cm, every liter of described strong bud medium contains: 6-BA1.0mg, NAA0.1mg, sucrose 40g, agar 6g, all the other are the MS medium, pH5.6, and its compound method is that above-mentioned substance is mixed, adjust pH, then high-temperature sterilization is standby.
2.4 culture of rootage: choosing is highly the plantlet of 3-4cm, the root induction in root media of transferring, and under 28 ℃ of conditions, luminous intensity is 1500lx.After 10 days, the seedling base section dissolves the former base of white root, can grow to the 5-7cm Banana Seedlings after 30 days, every liter of described root media contains NAA0.2mg, inositol 100mg, sucrose 40g, agar 6g, active carbon 1g, all the other are the MS medium, pH5.6, its compound method is that above-mentioned substance is mixed, adjust pH, then high-temperature sterilization is standby.
2.5 hardening and transplanting: choose well developed root system and healthy and strong Banana Seedlings, take out afterwash root medium, in greenhouse casting bed or Nutrition Soil, domestication, after 2 months, can be used as and transplants seedling replanting to field, then gives suitable rich water quality management and get final product.
Claims (2)
1. a new method that improves banana tissue-culturing rapid propagation coefficient, is characterized in that, comprises the following steps:
Choosing with bud of a, explant induced: get the suction bud of healthy banana plant base portion, clean, ream and inhale bud top layer and leaf sheath, leave and take the bastem section of inhaling, sterilization treatment, then peel off and inhale bud surface leaf sheath, leave and take the bastem section of inhaling, be placed on inducing culture, 28 ℃ of dark culturing, inhale sprout tuber until grow, and every liter of described inducing culture contains: 6-benzyl aminopurine 3-6mg, methyl α-naphthyl acetate 0.1-0.3mg, sucrose 30g, agar 6g, all the other are the MS medium, pH5.6-6.0;
The differentiation of b, bud and cultivation: will inhale the sprout tuber stripping and slicing, and be placed in differential medium and carry out the light cultivation, luminous intensity is 1500lx, 28 ℃, to the healthy indefinite bud of differentiation generation, indefinite bud is inoculated on differential medium, in luminous intensity, is 1500lx, 28 ℃ are carried out the subculture cultivation, subculture is cultivated and can be carried out some generations, obtains Multiple Buds, and every liter of described differential medium contains: 6-benzyl aminopurine 3-6mg, methyl α-naphthyl acetate 0.1-0.3mg, sucrose 35g, agar 6g, all the other are the MS medium, pH5.6-6.0;
C. the strong bud of Multiple Buds is processed: after Multiple Buds is divided into to little Cong or simple bud, put into strong bud medium, under 28 ℃ of conditions, luminous intensity is 1500lx, grow up to plantlet, every liter of described strong bud medium contains: 6-benzyl aminopurine 0.5-1mg, methyl α-naphthyl acetate 0.1mg, sucrose 40g, agar 6g, and all the other are the MS medium, pH5.6-6.0;
D, culture of rootage: choosing is highly the plantlet of 3～4cm, the root induction in root media of transferring, under 28 ℃ of conditions, luminous intensity is 1500lx, cultivate until grow up to Banana Seedlings, every liter of described root media contains methyl α-naphthyl acetate 0.1-0.2mg, inositol 50-100mg, sucrose 40g, agar 6g, active carbon 1g, and all the other are the MS medium, pH5.6-6.0;
F, hardening and transplanting: choose well developed root system and healthy and strong Banana Seedlings, take out afterwash root medium, in greenhouse casting bed or Nutrition Soil, after domestication, can be used as and transplant seedling.
2. the new method of raising banana tissue-culturing rapid propagation coefficient according to claim 1, it is characterized in that, reaming in step a inhaled bud top layer and leaf sheath, leave and take the bastem section of inhaling, sterilization treatment, peel off again and inhale bud surface leaf sheath, leaving and taking suction bastem section is to ream to inhale bud top layer and leaf sheath, leave and take the suction bastem section of 5cm, clean with volume fraction 75% alcohol water blend, and soak 10 minutes with mass fraction 0.1%HgCl2 solution, through aseptic water washing 4-5 time, blot surface moisture with aseptic filter paper again, finally leave and take the suction bastem section of 2cm, and peel off layer by layer and inhale bud surface leaf sheath.
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Cited By (9)
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| CN103960018A (en) * | 2014-05-14 | 2014-08-06 | 中国热带农业科学院海口实验站 | Method for increasing survival rate of transplanting banana temporarily-planted seedlings to big field |
| CN104982285A (en) * | 2015-06-17 | 2015-10-21 | 广东省农业科学院果树研究所 | Method for rapid propagation of banana seedlings based on suckers |
| CN105191789A (en) * | 2015-07-15 | 2015-12-30 | 航天神舟生物科技集团有限公司 | Purification and rejuvenation method for test-tube plantlets in banana tissue culture process |
| CN106688887A (en) * | 2016-12-08 | 2017-05-24 | 广东省农业科学院果树研究所 | Fenza No.1 dwarf banana tissue culture rapid propagation method |
| CN107027629A (en) * | 2017-05-04 | 2017-08-11 | 北海康维生态农业科技有限公司 | A kind of method for tissue culture for improving banana survival rate |
| CN107494278A (en) * | 2017-10-16 | 2017-12-22 | 李操 | A kind of quick-breeding method of Saigon any of several broadleaf plants tissue cultural seedlings of free |
| CN109247236A (en) * | 2018-11-09 | 2019-01-22 | 广西壮族自治区农业科学院园艺研究所 | More simplified sterilization method in a kind of banana tissue culture |
| CN113142055A (en) * | 2021-04-29 | 2021-07-23 | 广西壮族自治区农业科学院 | In-vitro proliferation preservation method for banana germplasm resources |
| CN115956505A (en) * | 2023-01-31 | 2023-04-14 | 高州市石生源生物科技发展有限公司 | Cultivation method of banana seedlings |
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| CN103960018A (en) * | 2014-05-14 | 2014-08-06 | 中国热带农业科学院海口实验站 | Method for increasing survival rate of transplanting banana temporarily-planted seedlings to big field |
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| CN104982285B (en) * | 2015-06-17 | 2017-06-06 | 广东省农业科学院果树研究所 | Based on the Banana Seedlings rapid propagation method for inhaling bud |
| CN105191789A (en) * | 2015-07-15 | 2015-12-30 | 航天神舟生物科技集团有限公司 | Purification and rejuvenation method for test-tube plantlets in banana tissue culture process |
| CN106688887A (en) * | 2016-12-08 | 2017-05-24 | 广东省农业科学院果树研究所 | Fenza No.1 dwarf banana tissue culture rapid propagation method |
| CN106688887B (en) * | 2016-12-08 | 2019-08-20 | 广东省农业科学院果树研究所 | The tissue culture and rapid propagation method of miscellaneous No. 1 dwarf banana of powder |
| CN107027629A (en) * | 2017-05-04 | 2017-08-11 | 北海康维生态农业科技有限公司 | A kind of method for tissue culture for improving banana survival rate |
| CN107494278A (en) * | 2017-10-16 | 2017-12-22 | 李操 | A kind of quick-breeding method of Saigon any of several broadleaf plants tissue cultural seedlings of free |
| CN109247236A (en) * | 2018-11-09 | 2019-01-22 | 广西壮族自治区农业科学院园艺研究所 | More simplified sterilization method in a kind of banana tissue culture |
| CN113142055A (en) * | 2021-04-29 | 2021-07-23 | 广西壮族自治区农业科学院 | In-vitro proliferation preservation method for banana germplasm resources |
| CN115956505A (en) * | 2023-01-31 | 2023-04-14 | 高州市石生源生物科技发展有限公司 | Cultivation method of banana seedlings |
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Application publication date: 20131218 |